Visscher M B, Lee Y C
Proc Natl Acad Sci U S A. 1976 May;73(5):1754-6. doi: 10.1073/pnas.73.5.1754.
Methods have been developed for the measurement of the Ca and Mg contents of subcellular particulates, separated by nonpolar liquid density gradient ultracentrifugation. After rapid deep freezing and removal of water by freeze-drying the frozen crushed tissue, homogenization and sonification a nonpolar liquid were carried out. Application of this technique has yielded evidence that the relative amounts of Mg and Ca are greatly different in the several subcellular particulates of normal heart muscle separated by density gradient centrifugation. It also appears that the comparative specific activity of Ca, after loading the perfusates with 45Ca, is very different in the several density fractions of the normal heart. The density site of greatest loading with 45Ca is changed in Ca2+-lack arrest and is very greatly altered by the induction of paradoxical Ca contracture. The method of ion-immobilization separation for water-free particulates of different densities in nonpolar liquids appears to be applicable to analysis for various constituents of other biological materials.
已开发出测量通过非极性液体密度梯度超速离心分离的亚细胞颗粒中钙和镁含量的方法。将冷冻粉碎的组织快速深度冷冻并通过冷冻干燥除去水分后，进行均质化并在非极性液体中进行超声处理。应用该技术已获得证据表明，通过密度梯度离心分离的正常心肌的几种亚细胞颗粒中镁和钙的相对含量差异很大。同样似乎是，在用45Ca加载灌注液后，正常心脏的几个密度级分中钙的比较比活性差异很大。45Ca最大负载的密度位点在缺钙性停搏时发生变化，并因矛盾性钙挛缩的诱导而发生很大改变。用于在非极性液体中分离不同密度的无水颗粒的离子固定化分离方法似乎适用于分析其他生物材料的各种成分。
